本研究探討以基因重組大腸桿菌E.coli JM109 生產肌酸酵素，此菌所殖入之載體PQE-51-PSCR01 內包含Pseudomonas putida NTU-8的肌酸酵素基因以及於其前端插入一段來自於Aeromonas hydrophila幾丁質分解酵素的信號序列，故所表現的胞內酵素可被分泌至細胞間質。將此基因重組菌於每升添加3.0 克葡萄糖之Luria-Bertani 培養液進行搖瓶培養，並在適當時間以IPTG 誘導，發現有促進菌體生長的現象。推測可能原因為誘導後，加速某些特定胺基酸之利用效率所致。於是再於R 培養液中添加不同胺基酸進行培養，結果發現添加蘇胺酸、精胺酸、組胺酸或蛋胺酸對於菌體的生長和肌酸酵素的生產皆有幫助。因此再於R 培養液中添加蘇胺酸、精胺酸及蛋胺酸三種胺基酸，並以回應曲面配合中心混成法尋找生產肌酸酵素之最佳條件，其結果似乎可觀察到高蘇胺酸濃度及低蛋胺酸濃度低較有利於肌酸酵素的生產

The ceatinase was produced from the gene of Pseudomonas putida NTU-8 expressed by a recombinant Escherichia coli JM109. This expression system was constructed by using a pQE-51-pSCR01 vector fused with a chitinase signal peptide, with which the gene product can excrete to the periplasmic space . Cell growth enchancement was found by incubating the recombinant E. coli in Luria-Bertani medium when an inducer of isopropyl-β-D-thiogalactopyranoside (IPTG) was added at an appropriate time. Such behavior might be attributed to the activated usage of certain amino acids. Similar behaviors of increasing cellgrowth and creatinase activing were obtained when adding threonine 、arginine 、histidine and methionine in the R medium. Therefore a Response Surface Method (RSM) with Central Composite Design (CCD) where threonine 、arginine and methionine were as the designed valuables were employed to find the optimal condition for creatinase production. The results indicated that the higher of threonine concentration and the lower of methionine concentration were, the better was the producivity for creatinase

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